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c-kit protein, a transmembrane kinase: identification in tissues and characterization
1Laboratory of Molecular Oncology, Memorial Sloan-Kettering Cancer Center, New York, New York.
Molecular and Cellular Biology
|November 1, 1988
Summary
Researchers identified the c-kit protein, a transmembrane kinase, in cat brain tissue using specific antibodies. This protein exhibits autophosphorylating activity and has complex carbohydrate structures, with variants found in mouse tissues.
Area of Science:
- Molecular Biology
- Oncology
- Neuroscience
Background:
- The proto-oncogene c-kit encodes a transmembrane kinase.
- c-kit is related to growth factor receptors and the immunoglobulin superfamily.
- Understanding c-kit's structure and function is crucial in cancer research.
Purpose of the Study:
- To identify and characterize the c-kit protein in cat brain tissue.
- To investigate the enzymatic activity of the c-kit protein.
- To explore variations of c-kit protein in different mammalian tissues.
Main Methods:
- Preparation of antibodies specific for the P80 gag-kit protein kinase domain.
- Immune complex kinase assays to detect autophosphorylating activity.
- Wheat germ agglutinin affinity chromatography for partial purification.
- Endoglycosidase digestion to analyze N-linked carbohydrates.
Main Results:
- A 145-kDa glycoprotein identified as c-kit was detected in cat brain.
- The c-kit protein displayed tyrosine-specific autophosphorylating activity.
- N-linked carbohydrates indicated hybrid, complex, and high-mannose structures.
- Cross-reactivity with murine c-kit revealed tissue-specific variants (145 kDa in brain, 160 kDa in spleen, 150 kDa in testis).
Conclusions:
- The study successfully identified and characterized the c-kit protein in feline brain.
- c-kit possesses autophosphorylating kinase activity crucial for its function.
- Tissue-specific variants of c-kit exist in mice, suggesting differential regulation or function.